JP2011004755A5 - - Google Patents

Claims (32)

Recombinant mutant Vaccinia Ankara (MVA) virus comprising at least two homologous foreign sequences with at least 50% identity, each of which is inserted at a different insertion site in the viral genome . A recombinant MVA virus comprising at least two homologous foreign sequences with at least 60% identity. The recombinant MVA virus according to claim 1 or 2, wherein the sequence has 65 to 75% identity. The recombinant MVA virus according to any one of claims 1 to 3, wherein the sequence is a homologous gene.   The recombinant MVA virus according to claim 4, wherein the homologous gene is derived from a flavivirus.   The recombinant MVA virus according to claim 5, wherein the flavivirus is a dengue virus.   The recombinant MVA virus according to claim 5 or 6, wherein the gene is at least two homologous genes derived from at least two different serotypes of the virus.   The recombinant MVA virus according to any one of claims 5 to 7, wherein the gene is at least two PrM genes.   The recombinant MVA virus according to any one of claims 5 to 8, wherein the gene is four PrM genes. The recombinant MVA virus according to any one of claims 1 to 3, wherein the homologous foreign sequences are identical. 11. The recombinant MVA virus according to claim 10, wherein the homologous foreign sequence is a promoter. The recombinant MVA virus according to any one of claims 4 to 9 and 11, wherein the genes are under the transcriptional control of the same promoter. The recombinant MVA virus according to claim 12 , wherein each of the homologous genes is under transcriptional control of vaccinia virus early / late promoter p7.5. The recombinant MVA virus according to any one of claims 1 to 13 , wherein the MVA is replication-defective or non-replicatable in mammalian cells including human cells. The recombinant MVA virus according to any one of claims 1 to 14 , wherein the sequence is inserted into a natural deletion site and / or intergenic region of the poxvirus genome. The recombinant MVA virus according to any one of claims 1 to 15 , as a medicament or a vaccine. A vaccine comprising the recombinant MVA virus according to any one of claims 1 to 15 . A pharmaceutical composition comprising the recombinant MVA virus according to any one of claims 1 to 15 and a pharmaceutically acceptable carrier, diluent, adjuvant and / or additive. 17. A recombinant MVA virus according to any one of claims 1 to 16 for influencing the immune response in living animals, including humans. 18. A vaccine according to claim 17 for influencing the immune response in living animals, including humans. 19. A composition according to claim 18 for influencing the immune response in living animals, including humans. Use of the recombinant MVA virus according to any one of claims 1 to 15 for the manufacture of a medicament. 23. Use according to claim 22 for the manufacture of a medicament for influencing the immune response in living animals, including humans. A cell comprising the recombinant MVA virus according to any one of claims 1 to 15 . A method for producing a recombinant MVA virus according to any one of claims 1 to 15 , comprising
-Infecting cells with MVA virus;
Sequence to be introduced into the genome of -MVA viruses, and the first vector construct comprising the genomic sequence of the MVA virus which have the ability to direct the incorporation of the insert to be arranged to the insertion site of the MVA viral genome transfected into the infected cells The process of
-Identifying , isolating and optionally purifying the produced recombinant MVA virus;
-Using the recombinant MVA virus obtained in the previous step for infection of the cells and another sequence to be introduced into the MVA virus genome, said sequence being homologous to the sequence of the first vector construct Repeating the above steps by using an additional vector construct comprising:
Including the above method. Two or more vector constructs, each construct comprising a sequence, the sequences contained in different vectors are homologous sequences having at least 50% identity, and each sequence contains a corresponding sequence to the MVA genome The vector construct, flanked by MVA viral DNA sequences having the ability to direct the integration of homologous sequences; and
-Means for identifying and / or selecting a recombinant MVA virus in which said homologous sequence is integrated in the genome;
Including kit. The each homologous sequence, MVA virus DNA sequence having the ability to direct the integration of said homologous sequence of the vector construct into different insertion sites of MVA virus genomes are adjacent, kit of claim 26. A polynucleotide derived from or homologous to the recombinant MVA virus genome of the recombinant MVA virus according to any one of claims 1 to 15 , comprising at least two homologous sequences and , A polynucleotide comprising at least part of the sequence of the MVA virus genome. A method for detecting a cell infected with the recombinant MVA virus according to any one of claims 1 to 15 , comprising administering the polynucleotide of claim 28 to the cell. 30. A method of identifying a recombinant MVA virus according to any one of claims 1-15 , comprising administering to the virus the polynucleotide of claim 28 . A method for detecting a cell infected with the recombinant MVA virus according to any one of claims 4 to 15 , wherein the cell is selectively selected from a homologous gene and / or a flanking sequence related to the insertion site of the gene. Contacting with a DNA primer to be amplified. A method for identifying the recombinant MVA virus according to any one of claims 4 to 15 , wherein the cell selectively amplifies a homologous gene and / or a flanking sequence related to an insertion site of the gene. The above method comprising contacting with a primer.